In vivo tracking of bone marrow stromal cells transplanted into mice cerebral infarct by fluorescence optical imaging
In vivo tracking of bone marrow stromal cells transplanted into mice cerebral infarct by fluorescence optical imaging
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DOI：
10.1016/j.brainresprot.2004.04.004
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发表时间：
2004-08-01
期刊：
影响因子：
--
通讯作者：
Iwasaki, Y
中科院分区：
文献类型：
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作者：
Shichinohe, H;Kuroda, S;Iwasaki, Y
最近的实验研究表明，骨髓基质细胞（BMSC）移植到各种神经系统疾病的动物模型中可以改善神经功能缺损，但确切的机制仍不清楚。在这项研究中，我们开发了一种新的体内荧光光学成像方案，以顺序跟踪移植到遭受脑梗塞的活体动物的大脑中。小鼠 BMSC 是从表达绿色荧光蛋白（BMSC-GFP）的转基因小鼠中收获的。它们被立体定向移植到缺血 7 天后进行永久性大脑中动脉闭塞的小鼠的同侧纹状体中（n = 12）。移植后12周内，暴露颅骨，使用体内荧光光学显微镜连续观察脑表面发出的绿色荧光。结果，在所有动物移植后4-12周，在同侧顶叶区域检测到区域绿色荧光，并且随着时间的推移变得更加明显。通过头骨获得的图像与通过减薄或去除头骨获得的图像非常相似。免疫组织化学评估显示，移植的细胞向缺血边界区迁移并表达神经元或星形细胞标记，支持荧光光学图像的发现。移植到活体动物大脑中的 BMSC 的连续可视化对于监测移植细胞的迁移、生长和分化具有重要意义，从而探索干细胞移植治疗各种神经系统疾病的命运和安全性。 (C) 2004 Elsevier B.V. 保留所有权利。
Recent experimental studies have indicated that bone marrow stromal cells (BMSC) improve neurological deficits when transplanted into the animal models of various neurological disorders, although precise mechanism still remains unclear. In this study, we developed a new in vivo fluorescence optical imaging protocol to sequentially track the transplanted into the brain of the living animals subjected to cerebral infarct. Mice BMSC were harvested from transgenic mice expressing green fluorescent protein (BMSC-GFP). They were stereotactically transplanted into the ipsilateral striatum of mice subjected to permanent middle cerebral artery occlusion after 7 days of ischemia (n = 12). During 12 weeks after transplantation, the skull was exposed and the green fluorescence emitted from the brain surface was sequentially observed, using in vivo fluorescence optical microscopy. As the results, regional green fluorescence was detected in the ipsilateral parietal region 4-12 weeks after transplantation in all animals and became more apparent over the time. The images obtained through the skull were very similar to those acquired by thinning or removing the skull. Immunohistochemistry evaluation revealed that the transplanted cells migrated towards the ischemic boundary zone and expressed the neuronal or astrocytic marker, supporting the findings on fluorescence optical images. Sequential visualization of the BMSC transplanted into the brain of living animals would be valuable for monitoring the migration, growth and differentiation of the transplanted cells to explore the fate and safety of stem cell transplantation for various neurological disorders. (C) 2004 Elsevier B.V. All rights reserved.